Rumen Bacteria Medium

[所属分类:培养基配方] [发布时间:2021-8-6] [发布人:网站管理员2] [阅读次数:] [返回]

Rumen Bacteria Medium

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per 1001.0mL:

Na 2 CO 3  .........................................................................................4.0g
Trypticase™..................................................................................2.0g
Yeast extract..................................................................................0.5g
K 2 HPO 4 .........................................................................................0.3g
Hemin ........................................................................................1.0mg
Resazurin...................................................................................1.0mg
Minerals solution.....................................................................38.0mL
Carbohydrate solution..............................................................20.0mL
L -Cysteine·HCl·H 2 O solution..................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Volatile fatty acid mixture .........................................................3.1mL
pH 6.7 ± 0.2 at 25°C
Minerals Solution:
Composition per liter:
NaCl............................................................................................12.0g
KH 2 PO 4 .........................................................................................6.0g
(NH 4 ) 2 SO 4 ....................................................................................6.0g
MgSO 4 ·7H 2 O................................................................................2.5g
CaCl 2 ·2H 2 O ..................................................................................1.6g
Preparation of Minerals Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
L -Cysteine·HCl·H 2 O Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O...................................................................0.25g
Preparation of  L -Cysteine·HCl·H2O Solution: Add  L -cyste-
ine·HCl·H 2 O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Sparge with 100% CO 2 . Autoclave for 15 min at 15
psi pressure–121°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.25g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% CO 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Carbohydrate Solution:
Composition per 20.0mL:
Glucose.........................................................................................0.5g
Cellobiose.....................................................................................0.5g
Glycerol ........................................................................................0.5g
Maltose .........................................................................................0.5g
Starch, soluble...............................................................................0.5g
Preparation of Carbohydrate Solution : Add components to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Sparge under 100% CO 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Volatile Fatty Acid Mixture:
Composition per 7.75mL:
Acetic acid...............................................................................4.25mL
Propionic acid..........................................................................1.50mL
Butyric acid................................................................................1.0mL
DL -2-Methyl butyric acid.........................................................0.25mL
iso-Butyric acid........................................................................0.25mL
iso-Valeric acid........................................................................0.25mL
n-Valeric acid...........................................................................0.25mL
Preparation of Volatile Fatty Acid Mixture: Combine compo-
nents. Mix thoroughly.
Preparation of Medium: Prepare and dispense medium under
100% CO 2 . Add components, except carbohydrate solution, Na 2 CO 3  ,  L -
cysteine·HCl·H 2 O solution, and Na 2 S·9H 2 O solution, to distilled/deion-
ized water and bring volume to 960.0mL Mix thoroughly. Gently heat
and bring to boiling. Continue boiling for 5 min. Cool to room tempera-
ture while sparging with 100% CO 2 . Add Na 2 CO 3 . Continue sparging
with 100% CO 2 until pH reaches 6.8. Distribute into rubber-stoppered
tubes under 100% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Aseptically and anaerobically add 20.0mL of sterile carbohydrate solu-
tion, 10.0mL of sterile  L -cysteine·HCl·H 2 O solution, and 10.0mL of ster-
ile Na 2 S·9H 2 O solution or, using a syringe, inject the appropriate amount
of sterile carbohydrate solution, sterile Na 2 S·9H 2 O solution, and sterile
L -cysteine·HCl·H 2 O solution into individual tubes containing medium.
Use: For the cultivation and maintenance of Anaerovibrio glycerini,
Anaerovibrio lipolytica, Butyrivibrio fibrisolvens, Lachnospira multi-
parus, Succinimonas amylolytica, and Succinivibrio dextrinosolvens.